Exposing synthetic lethal vulnerabilities in EBV-positive AIDS-NHL through novel replication dependency factors

通过新型复制依赖性因子揭示 EBV 阳性 AIDS-NHL 的综合致命脆弱性

基本信息

批准号：
10700376
负责人：
SUMITA BHADURI-MCINTOSH
金额：
$ 61.18万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-04 至 2028-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10700376
关键词：
AIDS-Related Lymphoma Acquired Immunodeficiency Syndrome Address B-Cell Activation B-Cell Lymphomas B-Lymphocytes Biopsy Blast Cell Blood Burkitt Lymphoma Cancer Etiology Cell Cycle Cell Death Cell Line Cell Nucleus Cell Proliferation Cells Cessation of life Complex DNA DNA biosynthesis DNA replication fork Dependence Development Disease Ensure Epstein Barr Virus lymphoma cell Epstein-Barr Virus-Related Lymphoma Event Genes Genetic Genome Goals HIV HIV/AIDS Human Herpesvirus 4 Immune Immunocompromised Host Immunologic Surveillance Impairment Individual Infection Influentials Investigation Job&apos s Syndrome Knowledge Link Lymphoma Lymphoma cell MCM7 gene Malignant Neoplasms Mass Spectrum Analysis Mechanics Modeling Morbidity - disease rate Mutation Non-Hodgkin&apos s Lymphoma Nucleotides Oncoproteins Pathway interactions Patients Persons Phase Predisposition Proliferating Property Proteins Proteome Proto-Oncogenes Public Health Risk Role S phase STAT3 gene Stress Testing Therapeutic Tonsil Trans-Activators Transcript Translating Transplant Recipients Up-Regulation Viral Virus Virus Latency Zinc Fingers antiretroviral therapy cancer cell cancer specimen resource cell transformation chemotherapy clinically relevant experience gammaherpesvirus genome-wide helicase immunosuppressed improved outcome in vivo infected B cell innovation large cell Diffuse non-Hodgkin&apos s lymphoma lymphoblastoid cell line mortality new therapeutic target novel oral pathogen recruit replication stress transforming virus tumor microenvironment

项目摘要

PROJECT SUMMARY Virus-associated lymphomas cause significant morbidity and mortality in HIV-infected individuals – indeed, the oral pathogen Epstein-Barr virus (EBV) contributes to up to 90% of diffuse large B-cell lymphomas (DLBCL) and 40% of Burkitt lymphomas (BL). Although combined antiretroviral therapy (cART) and chemotherapy have improved outcomes for AIDS lymphomas, challenges remain particularly with virus-associated AIDS lymphomas, prompting efforts to better understand virus-associated factors and pathways. In particular, EBV-driven cellular genome replication which is essential to lymphoma proliferation remains underexplored. Upon infection of B cells, EBV drives host DNA replication which is essential for establishment of viral latency as well as proliferation of cancer cells. However, such viral oncoprotein-driven DNA replication is plagued with physical and functional obstacles, resulting in replication stress. Such replication stress is a barrier to cancer. And yet, how EBV-cancer cells overcome such stress at replication forks to successfully proliferate is not well understood. In addressing this knowledge gap, we combined isolation of proteins on nascent DNA (iPOND) and mass spectrometry to discover novel fork proteins. This revealed a critical role for ZC3H18 (or ZC3) as a replication dependency factor that EBV upregulates to ensure host genome replication and lymphoma cell proliferation; notably, ZC3 had not been previously linked to DNA replication. Indeed, EBV+ DLBCL from AIDS patients have elevated ZC3 expression compared to EBV- lymphomas. An intrinsically disordered protein, ZC3 has the potential to concentrate a variety of proteins at replication forks. We find a direct interaction between ZC3 and MCM7 (a core component of the replicative helicase complex), further pointing to ZC3’s influential role in proliferation of EBV transformed cells. Importantly, ZC3’s partnership with other replication dependency factors exposes EBV-lymphoma cells to synthetic lethality – such therapies exploit the property that cancer cells tolerate perturbation of a single gene but succumb to co-disruption of multiple genetic events. In this application, we will test the hypothesis that EBV modulates the DNA replication machinery, ensuring proliferation of transformed cells in the face of replication stress and enhancing the potential for susceptibility to synthetic lethality. We will perform the following aims using ex vivo models and translate our results to patient- derived EBV+ AIDS lymphomas obtained from the AIDS and Cancer Specimen Resource (ACSR). Aim 1. Investigate how novel dependency factors unmask synthetic lethal vulnerabilities in EBV- transformed cells & Aim 2. Investigate mechanisms of ZC3 upregulation, replication machinery rewiring, and contribution of replication dependency factors to EBV+ AIDS lymphomas. These studies will identify mechanisms and generate new paradigms that reveal how an opportunistic virus modulates the host replication machinery to maintain the transformed state. Our long-term goal is to identify novel druggable targets that demonstrate synthetic lethality against EBV+ lymphomas in persons with HIV/AIDS.

项目概要病毒相关淋巴瘤会导致 HIV 感染者显着的发病率和死亡率——事实上，口腔病原体 Epstein-Barr 病毒 (EBV) 导致高达 90% 的弥漫性大 B 细胞淋巴瘤 (DLBCL) 和尽管联合抗逆转录病毒治疗 (cART) 和化疗已导致 40% 的伯基特淋巴瘤 (BL)。艾滋病淋巴瘤的治疗效果有所改善，但挑战仍然存在，特别是与病毒相关的艾滋病淋巴瘤，促使人们努力更好地了解病毒相关因素和途径，特别是 EBV 驱动的细胞。对于淋巴瘤增殖至关重要的基因组复制仍未得到充分研究。感染 B 细胞后，EBV 会驱动宿主 DNA 复制，这对于建立病毒潜伏期至关重要然而，这种病毒癌蛋白驱动的DNA复制受到困扰。身体和功能障碍，导致复制应激，这种复制应激是癌症的障碍。然而，EBV 癌细胞如何克服复制叉上的这种压力以成功增殖尚不清楚为了解决这一知识差距，我们结合了新生 DNA 上的蛋白质分离 (iPOND) 和质谱法发现新的叉蛋白，这揭示了 ZC3H18（或 ZC3）作为 EBV 上调以确保宿主基因组复制和淋巴瘤细胞的复制依赖因子值得注意的是，ZC3 以前并未与 AIDS 中的 EBV+ DLBCL 复制相关。与 EBV 淋巴瘤相比，患者的 ZC3 表达升高。ZC3 是一种本质上紊乱的蛋白质。具有将多种蛋白质集中在复制叉上的潜力，我们发现它们之间存在直接相互作用。 ZC3和MCM7（复制解旋酶复合物的核心成分），进一步表明ZC3的影响力作用重要的是，ZC3 与其他复制依赖性因子的合作。使 EBV 淋巴瘤细胞遭受合成致死——此类疗法利用了癌细胞耐受的特性单个基因的扰动，但会屈服于多个遗传事件的共同破坏。在此应用中，我们将测试 EBV 调节 DNA 复制机制的假设，确保转化细胞在面临复制应激时增殖并增强易感性的潜力我们将使用离体模型实现以下目标，并将我们的结果转化为患者- 源自艾滋病和癌症标本资源 (ACSR) 的 EBV+ 艾滋病淋巴瘤。目标 1. 研究新的依赖性因素如何揭示 EBV- 中的合成致命漏洞转化细胞和目标 2。研究 ZC3 上调、复制机器重新布线的机制，以及复制依赖因子对 EBV+ 艾滋病淋巴瘤的影响。这些研究将确定机制并产生新的范式，揭示机会性病毒如何调节宿主复制机制以维持转化状态。我们的长期目标是识别。新型药物靶标，证明对 HIV/AIDS 患者的 EBV+ 淋巴瘤具有合成致死性。